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MB Sample ID: SA006149

Local Sample ID:Middle exponential phase-P2
Subject ID:SU000135
Subject Type:Bacterial cells
Subject Species:Acidipropionibacterium acidipropionici
Taxonomy ID:1748
Genotype Strain:GCMCC1.2230/WSH1105
Species Group:Microorganism

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Subject:

Subject ID:SU000135
Subject Type:Bacterial cells
Subject Species:Acidipropionibacterium acidipropionici
Taxonomy ID:1748
Genotype Strain:GCMCC1.2230/WSH1105
Species Group:Microorganism

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
Middle exponential phase-P2SA006149FL001582Middle exponential phaseGrowth phase

Collection:

Collection ID:CO000119
Collection Summary:The cells were quenched by immediately adding a threefold volume of pre-chilled 60% (v/v) methanol solution containing 70 mM 4-(2-hydroxyethyl)-1-piperazine ethanesulphonic acid, and pelleted in a centrifuge (4,000 × g, ?4°C, 10 min)
Collection Protocol Filename:Protocol_Methods-G.docx
Sample Type:cell
Collection Time:Samples were collected at the middle exponential phase, late exponential phase, and the end of fermentation
Storage Conditions:-80°C
Collection Vials:50mL centrifuge tube
Storage Vials:50mL centrifuge tube
Collection Tube Temp:pre-chilled at -80°C
Additives:pre-chilled 60% (v/v) methanol solution containing 70 mM 4-(2-hydroxyethyl)-1-piperazine ethanesulphonic acid

Treatment:

Treatment ID:TR000137
Cell Storage:-80°C
Cell Growth Container:250 mL anaerobic jars
Cell Inoc Proc:inoculated at 1%
Cell Media:10 g/L yeast extract, 5 g/L tryptic soy broth, 1.5 g/L KH2PO4, 2.5 g/L K2HPO4
Cell Envir Cond:incubated at 32°C, anaerobic

Sample Preparation:

Sampleprep ID:SP000132
Sampleprep Summary:After quenching at –40°C, the cells were pelleted in a centrifuge (4,000 × g, ?4°C, 10 min). The pellets were washed with methanol solution and resuspended in 1 mL 35% perchloric acid to extract the metabolites from the cells. The mixture was frozen in liquid nitrogen and thawed three times. The supernatant was neutralized by adding K2CO3 solution with an initial concentration of 5 M, and an extract containing all metabolites was collected via centrifugation at 10,000 × g for 5 min at –4°C
Processing Method:Homogenization and solvent removal
Processing Storage Conditions:on ice
Extraction Method:35% perchloric acid and 5 M K2CO3
Extract Storage:-80°C
Cell Type:wildtype cells, genome shffled cells

Combined analysis:

Analysis ID AN000197
Analysis type MS
Chromatography type Reversed phase
Chromatography system LCMS-IT-TOF
Column Shim-Pack VP-ODS 150 L 2.0
MS Type ESI
MS instrument type IT-TOF
MS instrument name Shimadzu LCMS-IT-TOF
Ion Mode POSITIVE/NEGATIVE
Units Peak area

Chromatography:

Chromatography ID:CH000130
Chromatography Summary:LC-MS
Instrument Name:LCMS-IT-TOF
Column Name:Shim-Pack VP-ODS 150 L 2.0
Column Pressure:18 MPa
Column Temperature:40C
Flow Gradient:2% to 60% B for 15 min, 60% to 76% B for 15 min, and 76% to 2% B for 5 min (A:1 mM ammonium formate; B: 80% methanol (v/v) containing 1 mM ammonium formate)
Flow Rate:0.2 mL/min
Retention Time:35 min
Sample Injection:10 ?L
Solvent A:1 mM ammonium formate
Solvent B:80% methanol (v/v) containing 1 mM ammonium formate
Chromatography Type:Reversed phase

MS:

MS ID:MS000160
Analysis ID:AN000197
Instrument Name:Shimadzu LCMS-IT-TOF
Instrument Type:IT-TOF
MS Type:ESI
Ion Mode:POSITIVE/NEGATIVE
Collision Energy:0.4
Collision Gas:N2
Dry Gas Flow:1.5 L/min
Dry Gas Temp:350°C
Mass Accuracy:5ppm
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